Research status of additives related to platelet preservation / 中国输血杂志

Platelets play an important role in hemostasis, inflammation, tumor metastasis, wound healing and defense response, but its routine storage requires a specific temperature and the storage time is generally limited to 5 ~7 d, at the same time, platelet storage damage and bacterial breeding will limit its storage time. In addition, routine transfusion can cause serious adverse reactions such as non-hemolytic fever, allergies, circulatory overload, and transfusion-related acute lung injury. These limit the clinical application of platelet products. However, additives associated with platelet preservation can reduce the likelihood of their occurrence. Therefore, this article reviews the research status of additives related to platelet preservation in recent years, as well as improving platelet storage damage and improving platelet preservation characteristics.

Effect of trehalose on platelets stored at 4℃ in vitro / 中国输血杂志

【Objective】 To investigate the effect of trehalose added to M-Sol on platelet preservation at 4℃ in vitro. 【Methods】 Seven bags of ABO homotype concentrated platelets were randomly selected and divided into 7 groups according to storage temperature, preservation medium and trehalose concentration: 22℃+ PRP group, PRP group, M-Sol+ PRP group, M-Sol+ PRP+ 1 g/L trehalose group, M-Sol+ PRP+ 5 g/L trehalose group, M-Sol+ PRP+ 15 g/L trehalose group and M-Sol+ PRP+ 25 g/L trehalose group. PRP group and M-Sol preservation groups were stored at 4℃. Plt, PDW, MPV, maximum aggregation, CD62p positive rate, GLU and LAC concentration were detected on the 1st, 3rd, 5th and 7th day after preservation, and the changes of GLU and LAC concentration within 7 days were calculated. 【Results】 With the extension of preservation time, Plt decreased in all groups, and there was no significant difference among groups at the same time (P>0.05). PDW and MPV increased in all groups. When preserved to the 7th day, the PDW 10.22±0.43(fL) and MPV 8.74±0.40(fL) of M-Sol+ PRP+ 5 g/L trehalose group were the lowest, which was significantly different from that of 22℃+ PRP group (P0.05). During the preservation period, the maximum aggregation degree of each group decreased gradually. Except for the PRP group, the maximum aggregation degree of the M-Sol+ PRP+ 5g/L trehalose group was the highest(27.29±6.62), which was significantly higher than that of the 22℃+ PRP group on the 7th day after preservation (P<0.05). On the 7th day after preservation, the positive rate of CD62p in M-Sol+ PRP+ 5g/L trehalose group was the lowest(15.46±2.46), and there was significantly different compared with other groups (P<0.05). 【Conclusion】 Adding appropriate concentration of trehalose to M-Sol can inhibit platelet activation at 4℃ and reduce platelet storage lesion.

Comparative analysis of miRNA profiles of platelets treated and untreated with riboflavin and ultraviolet-B light / 中国输血杂志

【Objective】 To analyze the changes of microRNA (miRNA) expression profiles on day 1 and day 5 after storage with or without riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample), collected from voluntary donors, were split into two group after mixing and agitation. One was treated with riboflavin (final concentration 50 μmol/L) plus 6.24 J/mL UVB light(E group), and the other worked as a control group (C group) without any treatment. Both groups were subjected to agitated storage at (22±2) ℃ horizontally. The platelet concentrates were sampled on d1 and d5 (5mL) during storage, named as E1, E5, C1 and C5 groups, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between E and C groups were screened by using DEGseq and MA-plot analysis software, and GO function enrichment analysis and KEGG pathway enrichment analysis were further performed when the different expression between groups reached twofold and above. 【Results】 Compared with C1 group, 487 miRNAs with significantly different expression (P<0.01) were screened in E1 group, including 220 up-regulated miRNAs, such as miR-146a and let-7b, and 267 down regulated miRNAs, such as miR-7 and miR-1260. Compared with C5 group, 229 miRNAs with significantly different expression (P<0.01) were screened in E5, including 80 miRNAs with up-regulated expression, such as miR-423 and miR-378, and 149 down regulated miRNAs, such as miR-451 and miR-30.The target genes with differentially expressed miRNAs in E1 vs C1 groups and E5 vs C5 groups were similar in the numbers of enriched GO terms, including cell components, organelles, cell membrane and other cell structures, molecular functions such as adhesion, catalysis, molecular transformation, transportation, transcription factors and receptor activity, cell processing, metabolism, biological regulation, stress and other biological processes etc. Compared with E1 and C1 groups, E5 and C5 groups lacked of signal pathways related to environmental adaptation, translation and mucin synthesis, however, it increased inositol phosphate metabolism, phosphatidylinositol signaling system and chemokine signaling pathway. 【Conclusion】 The expression profiles of platelets miRNAs treated with VB2-PRT has changed significantly after storage for a period of time. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL induced by VB2-PRT.

Advances in applicaiton of platelets with cold storage at 4℃ and its storage lesion / 中国输血杂志

Platelet transfusion is one of the important therapeutic methods for clinical prevention of bleeding and hemostasis. At present, agitated storage at (22±2)℃used for platelets leads to risk of bacterial growth, which limits platelet shelf life and safety in transfusion. Cold storage at 4 ℃ can significantly promote the abovementioned drawbacks and present a better hemostatic effect. Platelets storage lesion(PSL) due to the cold storage, however, may lead to platelet activation following transfusion, thus reducing the effective survival rate in vivo. This paper reviews the research advances in PSL and the application of platelets stored at 4 ℃.

Expression analysis of miRNA profiles in apheresis platelets at the end of storage under riboflavin and ultraviolet-Blight treatment / 中国输血杂志

【Objective】 To investigate the changes of platelet microRNA (miRNA) expression profiles on d1 and d5 during storage with riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample) were collected from voluntary blood donors. After mixing and shaking, the samples was treated with riboflavin (final concentration 50 μmol/L) and 6.24J/mL UVB light for 8min, then split into two aliquots and agitated stored at (22±2) ℃. The concentrates were sampled (5mL) on d1 and d5, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between the two groups (at different storage periods) were screened by DEGseq and MA-plot analysis software. The miRNAs, reached more than 2 times different expression between groups, were considered significant different(P<0.01). The miRanda and TargetScan softwares were used to predict the target genes. Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on the target genes of significant differentially expressed miRNAs. The expression of miRNAs was verified by real-time fluorescence quantitative PCR (qRT-PCR). 【Results】 miRNA expression profile: compared with d1 platelets, there were 590 miRNAs with significantly different expression (P< 0.01) in d5 group, including 255 up-regulated miRNAs (such as miR-99b, miR-7) and 335 down regulated miRNAs (such as miR-451a, miR-19b). Among the 272 known miRNAs, 112 were up-regulated and 160 were down regulated. There were 318 new miRNAs sequences. The enriched GO terms of target genes of differentially expressed miRNAs in d5 and d1 groups included cell components, organelles, cell membranes and other cellular structures, molecular functions such as adhesion, catalysis, molecular conversion, transportation, transcription factor and receptor activity, and biological processes such as cell processing, metabolism, biological regulation and stress. The corresponding pathways in the top 10 of KEGG enrichment were mainly secretion, glucose metabolism, signal transduction, membrane transport, translation, environmental adaptation and other signal pathways. The six randomly selected differentially expressed miRNAs verified by qRT-PCR was consistent with those of DNB sequencing. 【Conclusion】 The expression profiles of platelets miRNAs has changed significantly between d1- and d5-storage under VB2-PRT treatment. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL underVB2-PRT treatment.

Mechanism of platelet activation via NF-κB signaling pathway causing human pulmonary microvascular endothelial cell injury / 中国输血杂志

【Objective】 To explore the role of NF-κB signaling pathway in the process of platelet storage on neutrophil-mediated human pulmonary microvascular endothelial cell injury, in order to further clarify the mechanism of platelet-mediated transfusion-related acute lung injury. 【Methods】 A co-culture system of human lung microvascular endothelial cells and neutrophils was established. A simple co-culture group and NF-κB inhibitor addition group were set up simultaneously, and then the platelet supernatant stored for 3 and 5 days were added. The expression levels of NF-κB pathway protein IκBα and cytokines (IL-6, IL-8, IL-1β and TNF-α) were detected, the cell death ratio (the number of dead cells/total number of cells) and the expression level of apoptotic protein Caspase 3 were calculated by Trypan blue staining. 【Results】 In the simple co-culture group with 5-day stored platelet supernatant added, the expression level of NF-κB pathway protein IκBα decreased significantly as the incubation time prolonged (simple co-culture 39 281.48±289.36 vs control 11 267.68±407.27, P0.05)when 5-day stored platelet supernatant was compared with 3-day stored one. The cell death rate(0.27±0.12 vs 3.33±0.31)and the expression level of apoptotic protein Caspase 3(17 821.11±611.55 vs 42 064.42±542.86)showed a significant increase(P0.05)as compared with the controls. Significant difference in the cell death rate of co-culture group with 5-day stored platelet supernatant added was observed as in comparison with the control (0.27±0.12 vs 1.26±0.11, P0.05). No significant difference was noticed in the expression level of apoptotic protein Caspase3(10 502.77±457.55 vs 10 424.16±471.53, P>0.05)when 5-day stored platelet supernatant was compared with 3-day stored one. 【Conclusion】 The NF-κB signaling pathway may be involved in storing platelets to regulate inflammation and apoptosis, thus affecting the process of neutrophil-mediated human pulmonary microvascular endothelial cell injury.

Evaluation of a domestic second generation platelet storage container / 대한수혈학회지

BACKGROUND: Polyvinyl (PVC) plastic container plasticized with di-(2-ethylhexyl) phthalate (DEHP) has been used for the storage of platelet concentrates for five days in Korea. Authors evaluated a second generation platelet storage container plasticized with tri (2-ethylhexyl) phthalate (TOTM) which was recently produced by Green Cross Medical Corp.(Korea). METHODS: 30 units of platelet concentrates were stored in TOTM-PVC container at 22'C in a flatbed agitator. Samples were taken at day 1,3,5, and 7 from the containers and tested for platelet count, MPV, PDW, pH, HCO3-,LDH, lactate, hypotonic shock response and beta-thromboglobulin (beta-TG). Electron microscopic examination was also performed. RESULTS: The number and functions of platelets were well preserved during storage. pH was maintained above 6.8 and any evidence for platelet activation was minimal. CONCLUSION: The TOTM-PVC second generation platelet storage container recently produced by the Green Cross Medical Corp.(Korea) was able to preserve platelets for at least five days without significant storage lesions.

Evaluation of Platelet Concentrates Stored in the Second Generation Platelet Storage Containers / 대한수혈학회지

BACKGROUND: Platelets can be stored for 3days at 22degrees C in conventional plastic bags plasticizer with di-(2-ethylhexyl)phthalate(DEHP). However, with such a short interval for storage, platelets could not be made easily available for thrombocytopenic patients. In vitro platelet function during 5 days of storage at 22degrees C was studied in a new plastic bag (second generation bag) which contained as plasticizer a tri (2-ethylhexyl)trimellitate and developed in Korea (Boin Medica Co.). METHODS: In vitro function (mean platelet volume, platelet distribution width, pH, platelet aggregation, platelet morphology and swirling phenomenon) was evaluated in 20 units of platelet concentrate at day 0, 2, 5 while mixing in 60 rpm platelet rotator at 22degrees C. RESULTS: At day 5, platelet count, mean platelet volume, platelet distribution width, pH, platelet aggregation, platelet morphology and swirling phenomenon were well maintained. pO2, pCO2 and HCO3- were 65.6mmHg, 43.4mmHg and 12.3mmol/L at day 5, respectively. CONCLUSION: The data indicate that the use of the new platelet storage container will permit satisfactory storage for at least 5 days at 22degrees C.

Effectiveness of additive solutions mixed with plasma on the storage of apheresis platelets / 中国输血杂志

6.0,and the expression of CD62P in groupⅠand Ⅱ(32% and 36%)was significantly higher than in control group(28%).However,when the three groups were stored until day 7,all in vitro parameters determined of PCs were better maintained in control group than in the other two.Conclusions The 5-day storage of PCs in additive solution with 50% or 20 % plasma is feasible in terms of the in vitro function in platelet count and pH,however,PCs storaged in additive solutions are more easily activated.